LOCUS       sequence-8951-ex        7846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7846)
  AUTHORS   Zhang Y, Nash L, Fisher AL.
  TITLE     A simplified, robust, and streamlined procedure for the production 
            of C. elegans transgenes via recombineering
  JOURNAL   BMC Dev Biol. 2008 Dec 30;8(1):119.
  PUBMED    19116030
REFERENCE   2  (bases 1 to 7846)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC Dev 
            Biol."; date: "2008-12-30"; volume: "8(1)"; pages: "119"
COMMENT     Sequence Label: ploxP unc-119
FEATURES             Location/Qualifiers
     source          1..7846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..51
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     36..52
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2009..2020)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    5832..5850
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     primer_bind     complement(5916..5933)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(join(6391..7182,7183..7251))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7014..7033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7252..7356)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7424..7442
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7480..7502)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7602..7621
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    7760..7778
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     protein_bind    join(7831..7846,1..18)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 tatgctatac gaagttatga attcgggccc cccctcgagg tcgacggtat cgataagctt
       61 cagtaaaaga agtagaattt tatagttttt tttctgtttg aaaaattctc cccatcaatg
      121 ttctttcaaa taaatacatc actaatgcaa agtattctat aacctcatat ctaaattctt
      181 caaaatctta acatatctta tcattgcttt aagtcaaacg taacattaaa aaaaatgttt
      241 tggaaaatgt gtcaagtctc tcaaaattca gttttttaaa ccactcctat agtcctatag
      301 tcctatagtt acccatgaaa tccttatata ttactgtaaa atgtttcaaa aaccattggc
      361 aaattgccag aactgaaaat ttccggcaaa ttggggaacc ggcaaattgc caatttgctg
      421 aatttgccgg aaaaacggta attgccgaaa gtttttgaca cgaaaatggc aaattgtggt
      481 tttaaaattt ttttttttgg aaatttcaga atttcaattt taatcggcaa aactgtaggc
      541 atcctaagaa tgttcctaca tctattttga aaagtaagcg aattctatga aaatgtctaa
      601 agaaaatggg gaaacaattt caaaaaggca cagtttcaag tgtttccgaa ttatactaaa
      661 tccctctaaa aacttccggc aaattgatat ccgtaaaaga gcaaatccgc atttttgccg
      721 aaaattaaaa tttccgacaa atcggcaaac cggcaatttg gcgaaatttg ccggaaggca
      781 attgccgccc acccctgttc cagaggttca aactggtagc aaagctcaaa atttctcaaa
      841 ttctccaatt tttttttgaa ttttggcagt gtaccaaaat gacattcagt catattggtt
      901 tattatagat ttatttagat aaaatcctaa atgattctac ctttaaagat gcccacttta
      961 aaagtaatga ctcaaacttc aaattgctct aagattctat tgaattacca tcttttcctc
     1021 tcattttctc tcactgtcta tttcatcaca aattcatccc tctctccctc tctttctctc
     1081 tccctctctc tctttctctt tgctcatcat ctgtcatttt gtccgttcct ctctctgcgc
     1141 cctcagcgtt ccccacactc tctcgcttct cttttcctag acgtcttctt ttttcatctt
     1201 cttcagccct ttttcgccat tttccatctc tgtcaatcat tacggacgac ccccattaat
     1261 ttttggatgt gctctgcgag caagagcaca cggaactcat ttattttttg ttggaatttc
     1321 tttttcattt tagttagtta aaaggttatt cagtgcaaaa attgattttt tgctcttttg
     1381 gtacattaga ggggtttgaa agttttccat aaaaagggtt tccggagtaa aaaatctaaa
     1441 aatttttaca aacatttcca aaaattttgg gaacatatct tttcaaaaac tcaaaaaaaa
     1501 acgcaaacat cataaattta atcaattaat tggatgcgtt caaatatttt aaaaatattt
     1561 ttgaattgca tatttcttta ctcataaaaa aattattaat gtttaagtaa aaaaaatatt
     1621 tcaggatttt aaacaaaaac tgtaaatccc ataaaacagt gttatttttt agttattttt
     1681 agtaattatt tgtgaattaa aaacacattt tttttcaaaa tattccttga ataaataaaa
     1741 ttgaaagtga attcaaaata tttcaccaat tttacaaaat ttgaaaattt gaaaaaatat
     1801 tttctaccaa attgtgctca aaaaatatta ttaagttccc aggaaattta tgatgaaaac
     1861 ttggaatatt tttggtaaaa ttttctcaaa aaattgaaag ttgtcaaaaa cacactaaca
     1921 ctatacctcg cggcatagaa aaaactggtg gccgaatttt ttaaaccaat ttaaattaaa
     1981 tatatttttg cactttgtac aaaaaaattt gaagttccca ggaaattcat acggaaatat
     2041 tttccagaac ttttataatt tttattttaa ttctcaaaat ttacaataaa aacttggaaa
     2101 cctgaaattt gcttaaagaa taaccctatc ccccacggcc tagaaaatac tggtggccga
     2161 attttttccg cagccacacc acctctaatc tccagagcac tctccaaaat tcccataatc
     2221 cccaaaattt tcagctacaa cagcatatat gaaggcagat ctacaacaac aatcgatcgc
     2281 acccggctcg gcaaccttcc cgtctcaggt gaggaagacg cagagaaatg agagagagag
     2341 cgtgtttatg cacgcagtat actaactcct gatatccaat ttcatatatc gatatgcttg
     2401 ctttcttttc ttctacgttg aaattgtttg aacttttctc aaaaaacgcg gattttagag
     2461 agacttagag ttgctccaaa taaatttaat ttgcaatttt gcacttacta ttagaatcta
     2521 cttttatagc gctaaaatga ccgaataaca cagtagattt taaaagaaaa ttttaaaaat
     2581 tatttttttg tgcagtcaaa tttcgctagt ttttgccact catttttgaa catttgtata
     2641 tgtgcctttc ggtggttcaa tcaggggtgg cacgtggtgt cagagtgttc catttcggtt
     2701 tgatctacgt tgatctacaa aaaatgcggg agaagagacg cagagttctc aactgatttc
     2761 acatggttat gaaccagggg tgttcggcaa atttgccgtt tgccgagctg ggcaaatttt
     2821 gagatttgcc gcacacccct tttatgaacg tgctgacgtc acatcttttt ggacaaagaa
     2881 ttcccgcatt ttttgtatat caaaccgtga tgggacagcc tggcaccacg tggggttggc
     2941 ggcaaatgat tttttccgac aaatcggcaa attggcaaat tgccggaatt taaagtttct
     3001 ggcaaattgt tgatttgccg aaaaaaaatt tcaaaaaggc acagttttaa gtgtttccgt
     3061 cttataaaat atttctctaa aaatttccgg aaaacggaaa attgccggaa atacaaagtt
     3121 tcgtcaaatc ggcaaacctg caatttgccg aacggcaact gccgccaaca cctggttccc
     3181 aaaaaagcgt gcactgtttg aatattaaaa aaaaaattca cacaaaaata ataaattgaa
     3241 ataggctaaa ttcacatcta tgtacaaaaa ttgcatgcca gcaccggtca attttctcaa
     3301 ttactttttg tctatttttg ccgaacataa tttgctgttt ccttggcaga atacagaaaa
     3361 ctcttttttt tttcagatgc cccgaccccc accggtaacc gaacaggcta taaccaccga
     3421 ggcggagctt ctcgcgaaaa atcaaattac accaaatgat gtgctagctc ttccaggaat
     3481 cactcaaggt aagtgaatat ttgctctatt gataaacgcg ccgtgtactc cacgtggaca
     3541 aacatatcaa attttcagga ttcctatgct ccccatcggc aaacgtctat aacattgagt
     3601 tcaccaaatt ccaaatccgt gacctcgaca cggaacacgt gctcttcgaa attgcaaaac
     3661 cggaaaatga gacggaagag aatctgcagg cgcaagccga atcggcaaga tatgtccgat
     3721 atcgatttgc gccgaatttt ctgaaattaa agacggtcgg cgcgacggtc gagttcaagg
     3781 tcggcgatgt gccgatcaca cattttcgaa tgattgaacg gcactttttc aaggatcgtt
     3841 tattgaaatg ctttgatttt gaatttggat tctgtatgcc aaattcgaga aacaactgtg
     3901 aacatatcta tgaatttcca caactttcac aacaactcag tgagtttgaa gcgggaaaaa
     3961 ttatttgagc tcattttgtc aatagagcga attttctagg tttttaaaaa taaaataaat
     4021 attgacaaaa agctgcccta cagtagttca aaatgtgaaa acttggagga aattcgagtt
     4081 ttctctgaga agtttttggc gggtaattca aattttctga gaaaaaaatt agccgcgaga
     4141 attcaaattt tctgagaaaa ttatttgacg ggaaattcaa aatttctgga aatgtattga
     4201 acatttgaga aattttgtta aaaactacat taaaccgata aaaaagttcg caaatttttt
     4261 ttaagtgctc atggtgcatc gaagagaaga attcttgcca aaactagccc atgaaattaa
     4321 aaattatcgc ggaaaaaaac tgactaattg gaaagtaaaa tgaaatttgc aaaaaaaaaa
     4381 tatgtttttt tgttgtaaaa ttaaaaaaaa aactgtatat tttcagtgga cgacatgatc
     4441 aataatccaa acgagacccg ctccgatagc ttctattttg tggagaataa gctcgtaatg
     4501 cacaataagg ccgactactc gtatgatgca taatttcccg ccgatggaat aaaactaata
     4561 atttttctcg caatttatag aaatcatgtg attttcctac cgctaaattt aaaatttcct
     4621 aatttttcac tatatttctc acaaaatgca ttccgaaaac tctcattttt cagcttttca
     4681 atacaaatcc caataaatga tttttcaaat ataatgttct caattgatat gttttatttg
     4741 aatgttggaa aaaaccataa aattaaaaaa aacttgaatt attcaaaaat aattcggccc
     4801 ttttacaaaa aattatcata aaaattaaat ttggaaaatc tgtgatttgt cgctttgccg
     4861 gaagtttcca attccggcaa ctaaccgatt tgccgattag ctagaaattt caattccgac
     4921 aatgtgccgc tttgccgatt tgccggaaat ttccaattcc ggcaattaac ggacttgccg
     4981 attagctaga aatttaactt tagataaaac ttctcaacaa aatcaaattg gaaagtcagc
     5041 gatttgccgc tttgccggaa gttttcagtt ccggcatcta accgatttgc cgatttgcca
     5101 aaaatgtcaa ttccgaaaaa ctgccgattt gccgctttgt cggaaatttc aattccgaca
     5161 aattgccgat ttgccgattt gccgaacatt ccaattccga ccatttgtcg atttgccgga
     5221 agttaccaat tccggcaact aacggatttg ccgatttgcc ggaaaaaaat tgtttgtcgc
     5281 caaccctgat ttaaatgact aatcatgttg caaaagataa ctgcttgcat caaaaatccg
     5341 atcattggca ttcccaacat cttcttcatc caaaacacca cccttttctt ccactaatca
     5401 tgaattcccg accaattgtt atcagtagga gaagcctcct tttatctcat tatcagcaat
     5461 tgccttcttc ttcttcttct ttgaatacca tttgaatgga gatatcattt acataaatct
     5521 gccgccgaga ttgtgaagca ggaggaaaag ctgaagatac tgatattata tgtatatttc
     5581 attattcttt ttccacaaaa ttttccgaca tttaattcca cttcactttc atcgtctctt
     5641 tttcccccat tgtcgccccc actcatttgt cttcttcttc aaatacgggt caatcacttt
     5701 ccgtttttta ttcatttttt gttatgttcg gcttaacaaa tttgaagtat tgttaaaatt
     5761 ctctagagtc gacctgcagg catgcaagct tgccaacgac tacgcactag ccaacaagag
     5821 cttcagggtt gagatgtgta taagagacag ctgtcttaat gaatcggcca acgcgcgggg
     5881 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5941 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6001 gaatcagggg ataacgcagg aaagaacatg gtgaaacctt aaaagctagc tttaaaagcc
     6061 ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat
     6121 ttatattaat tttattgttc aaacatgaga gcttagtacg ttagccatga gagcttagta
     6181 cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc
     6241 ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca
     6301 acaggttgaa ctgctgatct tcggatgcac aaattaaaaa tgaagtttta aatcaatcta
     6361 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6421 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6481 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6541 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6601 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6661 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6721 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6781 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6841 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6901 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6961 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7021 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7081 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7141 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7201 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7321 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7381 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7441 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7501 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     7561 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     7621 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     7681 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7741 ctcttcgcta ttacgacagc tgtctcttat acacatctca accatcatcg atgaattttc
     7801 tcgggtgttc tcgcatattg gctcgaattg ataacttcgt ataatg
//
